Squalene Bioproduction Engineering Service

Squalene is a high-value triterpene widely used in cosmetics for its moisturizing properties and in vaccines as an adjuvant. Traditionally, Squalene was sourced from shark liver oil, a method that shark extraction damages ecology and is unsustainable. While plant sources (like olive oil) are available, they suffer from low extraction amount and high purification costs. This drives the need for a highly efficient, sustainable, and ethical supply chain.

CD Biosynsis offers a synthetic biology service focused on establishing a high-titer microbial production route. Our core strategy involves modification of yeast terpenoid synthesis pathway (specifically the MVA pathway in Saccharomyces cerevisiae ) to maximize the flux of precursors (FPP) toward Squalene production. This is coupled with the overexpression of squalene synthase (ERG9) to create a strong metabolic sink, drawing maximum carbon flux towards the final product, while simultaneously blocking the downstream pathway (sterol synthesis) that consumes Squalene. This integrated approach aims to deliver a high-yield, pure, and environmentally sustainable Squalene product.

Pain Points

Establishing an ethical and cost-effective Squalene production route faces these critical limitations:

• Unsustainable Sourcing: Relying on shark liver oil leads to ecological damage and raises ethical concerns, prompting a global shift toward alternative sources.
• Low Titer in Bioproduction: In native hosts like yeast, Squalene is a metabolic intermediate quickly converted into sterols (like ergosterol) for cell membrane formation, leading to low accumulation.
• Pathway Bottlenecks: The upstream terpenoid pathway (MVA pathway) contains multiple feedback inhibition points and rate-limiting steps that restrict the supply of the precursor farnesyl pyrophosphate (FPP).
• Product Toxicity: High intracellular accumulation of lipids and terpenoids like Squalene can be toxic to the host yeast , limiting the maximum final yield.

A cost-effective solution must prevent the downstream consumption of Squalene and boost the upstream supply of its precursor.

Solutions

CD Biosynsis utilizes advanced synthetic biology and metabolic engineering to optimize Squalene production in yeast:

Modification of Yeast Terpenoid Synthesis Pathway

           

We overexpress rate-limiting enzymes in the MVA pathway (HMG-CoA reductase) and remove feedback inhibition to increase the supply of FPP.

Overexpression of Squalene Synthase (ERG9)

We overexpress the ERG9 gene and simultaneously knock out the downstream enzyme (ERG7, Squalene epoxidase) to prevent conversion to sterols, maximizing Squalene accumulation.

Host Tolerance Engineering

We modify lipid droplets and membrane composition to improve the host's tolerance to high intracellular Squalene concentrations, allowing for higher titers.

Balanced Cofactor Supply

Squalene synthesis requires NADPH. We optimize pathways like the pentose phosphate pathway (PPP) to ensure a constant supply of NADPH for the pathway to function efficiently.

This systematic approach focuses on overcoming the natural metabolic constraints to channel maximum carbon flux towards the final desired product, Squalene.

Advantages

Our Squalene engineering service is dedicated to pursuing the following production goals:

High Titer and Yield

Pathway engineering and sink creation lead to significantly higher Squalene accumulation per liter of culture than natural sources.

Sustainable and Ethical Sourcing

Microbial production provides an animal-free, renewable alternative , avoiding ecological damage from shark harvesting.

Reduced Purification Costs

High product specificity (blocking sterol synthesis) and concentration simplify downstream extraction and purification .

Established Host Platform

Utilizing S. cerevisiae (Baker's Yeast) provides a safe, robust, and well-understood industrial platform for fermentation.

High Purity Product

Precise genetic modification ensures the final product is primarily Squalene, minimizing unwanted byproducts .

We provide a biosynthetic platform aimed at maximizing the yield and minimizing the ethical/environmental footprint of Squalene production.

Process

Our Squalene strain engineering service follows a standardized, multi-stage research workflow:

• Upstream Pathway Enhancement: Overexpress rate-limiting enzymes (tHMG1) in the MVA pathway to increase the supply of the precursor FPP.
• Downstream Pathway Blockage: Knock out the squalene epoxidase gene (ERG1) and other sterol synthesis genes to prevent Squalene consumption.
• Squalene Synthase Overexpression: Introduce a strong promoter to overexpress Squalene Synthase (ERG9) to effectively convert all available FPP to Squalene.
• Cofactor and Tolerance Engineering: Engineer the host to increase NADPH availability and enhance the capacity of lipid bodies to store Squalene safely.
• Fermentation Performance Validation: Test the final engineered strain in fed-batch fermentation to assess Squalene titer, yield, and purity .
• Result Report Output: Compile a detailed Experimental Report including gene modification data, metabolic flux analysis, and fermentation metrics (yield, titer, and purity) , supporting commercial scale-up.

Technical communication is maintained throughout the process, focusing on timely feedback regarding yield and product accumulation.

Explore the potential for a high-performance, sustainable Squalene supply. CD Biosynsis provides customized strain engineering solutions:

• Detailed Metabolic Flux and Titer Analysis Report , demonstrating success in carbon redirection and titer increase.
• Consultation on primary extraction and downstream purification strategies optimized for yeast-produced Squalene.
• Experimental reports include complete raw data on carbon yield (g Squalene/g sugar) and lipid accumulation profile , essential for commercial application.